PMID- 10391953
OWN - NLM
STAT- MEDLINE
DCOM- 19990805
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 28
DP  - 1999 Jul 9
TI  - Characterization of DNA recognition by the human UV-damaged DNA-binding protein.
PG  - 20027-33
AB  - The UV-damaged DNA-binding (UV-DDB) protein is the major factor that binds DNA
      containing damage caused by UV radiation in mammalian cells. We have investigated
      the DNA recognition by this protein in vitro, using synthetic oligonucleotide
      duplexes and the protein purified from a HeLa cell extract. When a 32P-labeled
      30-mer duplex containing the (6-4) photoproduct at a single site was used as a
      probe, only a single complex was detected in an electrophoretic mobility shift
      assay. It was demonstrated by Western blotting that both of the subunits (p48 and
      p127) were present in this complex. Electrophoretic mobility shift assays using
      various duplexes showed that the UV-DDB protein formed a specific, high affinity 
      complex with the duplex containing an abasic site analog, in addition to the
      (6-4) photoproduct. By circular permutation analyses, these DNA duplexes were
      found to be bent at angles of 54 degrees and 57 degrees in the complexes with
      this protein. From the previously reported NMR studies and the fluorescence
      resonance energy transfer experiments in the present study, it can be concluded
      that the UV-DDB protein binds DNA that can be bent easily at the above angle.
FAU - Fujiwara, Y
AU  - Fujiwara Y
AD  - Department of Bioorganic Chemistry, Biomolecular Engineering Research Institute, 
      6-2-3 Furuedai, Suita, Osaka 565-0874, Japan.
FAU - Masutani, C
AU  - Masutani C
FAU - Mizukoshi, T
AU  - Mizukoshi T
FAU - Kondo, J
AU  - Kondo J
FAU - Hanaoka, F
AU  - Hanaoka F
FAU - Iwai, S
AU  - Iwai S
LA  - eng
PT  - Journal Article
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (DDB1 protein, human)
RN  - 0 (DNA Probes)
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (Oligodeoxyribonucleotides)
RN  - 9007-49-2 (DNA)
RN  - EC 4.1.99.3 (Deoxyribodipyrimidine Photo-Lyase)
SB  - IM
MH  - Base Sequence
MH  - DNA/*chemistry/radiation effects
MH  - *DNA Damage
MH  - DNA Probes/chemistry
MH  - DNA-Binding Proteins/*chemistry
MH  - Deoxyribodipyrimidine Photo-Lyase/chemistry
MH  - HeLa Cells
MH  - Humans
MH  - Molecular Sequence Data
MH  - Nucleic Acid Conformation
MH  - Oligodeoxyribonucleotides/chemistry
MH  - Substrate Specificity
EDAT- 1999/07/03 00:00
MHDA- 1999/07/03 00:01
CRDT- 1999/07/03 00:00
PHST- 1999/07/03 00:00 [pubmed]
PHST- 1999/07/03 00:01 [medline]
PHST- 1999/07/03 00:00 [entrez]
AID - 10.1074/jbc.274.28.20027 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Jul 9;274(28):20027-33. doi: 10.1074/jbc.274.28.20027.